1.Effects of PPAR-α silence on ET-1 induced myocardial hypertrophy and PI3K/Akt/GSK3β-NFATc4 signal pathway
Ruifang LI ; Kang LE ; Jie GAO ; Guoqing YANG ; Yingxia BAO ; Peiqing LIU
Chinese Journal of Pathophysiology 2009;25(12):2289-2294
AIM: To investigate the role and signal mechanism of PPAR-α in the pathogenesis of cardiac hypertrophy. METHODS: Small interfering RNA (siRNA) was applied to efficiently silence the gene expression of PPAR-α in cardiac myocytes. [~3H] leucine incorporation assay was performed to measure protein synthesis. Reverse transcription-polymerase chain reaction (RT-PCR) was used to analyze the mRNA level of atrial natriuretic factor (ANF) and PPAR-α. Western blotting analysis was performed to investigate the levels of phosphorylation of protein kinase B (PKB/Akt) and glycogen synthase kinase 3β (GSK3β). Immunofluorescence analysis was used to examine the cellular localization of NFATc4. RESULTS: (1)RSS304168 was the most efficient stealth RNAi duplex to specifically inhibit PPAR-α expression. (2)RSS304168 significantly potentiated the ET-1-induced cardiomyocyte hypertrophy and enhanced ET-1-induced protein synthesis and ANF mRNA expression in cardiomyocytes. Moreover, RSS304168 completely reversed the inhibitory effects of fenofibrate on ET-1-induced protein synthesis and ANF mRNA expression. (3)RSS304168 enhanced ET-1-induced phosphorylation of Akt at Ser473 and GSK3β at Ser9. The effects of ET-1 or ET-1 combined with RSS304168 on phosphorylation of Akt/GSK3β were completely blocked by LY294002, a PI3K specific inhibitor. Fenofibrate markedly inhibited ET-1-induced phosphorylation of Akt/GSK3β while RSS304168 abolished these effects of fenofibrate. (4)Fenofibrate prevented the nuclear translocation of NFATc4 induced by ET-1 while RSS304168 abolished this effect of fenofibrate. CONCLUSION: Activation of PPAR-α inhibits ET-1-induced cardiomyocyte hypertrophy through blocking Akt/GSK3β-NFATc4 signaling pathways.
2.Establishment of T-lymphocytes that express CD20scFv-IgGFc-CD28-? and CD20scFv-IgGFc and their killing activity of B-lymphoma cells
Yingxia TAN ; Kang YU ; Yongxian HU ; Shenghui ZHANG ; Shenmeng GAO ; Jianbo WU
Chinese Journal of Pathophysiology 2000;0(07):-
AIM:To investigate the target killing effect of T lymphocytes with chimeric CD20scFv gene on Daudi cells and the activation of T lymphocytes.METHODS:Two kinds of plasmids were transfected into retrovirus-packed PA317 cell lines.The supernatant was collected from successfully transfected PA317 culture and was used to infect peripheral blood T lymphocytes.After one-week screening with G418,the cells were used to kill Daudi and K562 cells.The positive rates of AnnexinⅤ in Daudi cells were measured at different times points respectively by flow cytometry.Meanwhile,the level of IL-2 and IFN-? were determined by ELISA.RESULTS:The Annexin V positive rate was significant higher in Daudi cells compared to control K562 cell lines at 24 h.No difference of AnnexinV in Daudi cells was observed in CD20 modification T lymphocyte groups.The secretions of IL-2 and IFN-? in CD20scFv-CD80-IgGFc-CD28-? gene modified T cells co-cultured with Daudi cells were dramatically higher than that in CD20scFv-IgGFc group at 72 h.CONCLUSION:① The two kinds of genetic modified specific T cells have no significant difference in inducing early apoptosis of Daudi cells.CD28-? can't affect Daudi cell early apoptosis at the CD20scFv target killing.② The increase in the secretions of IL-2 and IFN-? is more obvious in CD20scFv-IgGFc-CD28-? group,indicating that the self-activation takes place in CD3? and CD28 modified T cells without MHC restriction and then increases the activation and killing function of T cells.
3.Enzymatic removal of α-Gal antigen in porcine skin
Zhimin YUN ; Subo LI ; Xue ZHANG ; Yingxia TAN ; Shouping JI ; Hongwei GAO ; Feng GONG
Military Medical Sciences 2015;39(12):938-940
Objective To reduce immunogenicity of porcine skin by removingα-Gal epitopes expressed in cell surface and extracellular matrix using recombinant α-galactosidase produced by Bacteroides fragilis.Methods The porcine skin was harvested from healthy 2-month-old pigs without any skin disorders before being sterilized by iodine and 75%alcohol, respectively.Enzymatic removal of α-Gal antigen was followed by washing with PBS.The α-Gal antigen in the prepared porcine skin was measured with immunofluorostaining of cryosections and the residual enzyme was measured with a double-antibody sandwich ELISA method.Enzymatic removal procedures were optimized by detecting residual enzyme and the effi-cacy ofα-Gal removal under different enzymatic and washing conditions.Results Efficient enzymatic and washing methods were established to removeα-Gal antigen.Theα-Gal removal efficacy was above 90% and residual enzyme was undetect-able (αprescribed minimum ofα-galactosidase detection with indirect ELISA was 1 ng/ml) .Conclusion It is feasible to efficiently removeα-Gal antigen under these enzymatic and washing conditions, and a method of producing low-immunoge-nicity pig skin dressing for burn is established.
4.Chemical constituents from Aster sampsonii.
China Journal of Chinese Materia Medica 2010;35(17):2289-2292
From the whole plants of Aster sampsonii, ten compounds were isolated, and were characterized as (4alphaR, 4'alphaR, 5S, 5'S, 9alphaR, 9'alphaR)-4, 4', 4alpha, 4'alpha, 5, 5', 6, 6', 7, 7', 8, 8'-dodecahydro-3, 3', 4alpha, 4'alpha, 5, 5'-hexamethyl-2H, 2'H-9alpha, 9' alpha-binaphtho[2, 3-b]furan-2, 2'-dione(1), furanoligularenone(2), (4alphaR, 5S, 8alphaR)-4alpha, 5, 6, 7, 8, 8alpha-hexahydro-8alpha-hydroxy-3, 4alpha, 5-trimethylnaphtho[2, 3-b]furan-2(4H)-one(3), (4alphaR, 5S, 8alphaS)4alpha, 5, 6, 7, 8, 8alpha-hexahydro-8alpha-hydroxy-3, 4alpha, 5-trimethylnaphtho[2, 3-b]furan-2(4H)-one(4), methyl-1'-hydroxy-2', 6'-dimethoxy-4'-oxocyclohexanacetate(5), 1'-hydroxy-2'-methoxy-4'-oxocyclohexanacetate methyl(6), loliolide (7), jacaranone (8), beta-sitosterol(9), friedeline (10). The structures of all compounds were elucidated on the basis spectroscopic data, including IR, EI-MS, HR-ESI-MS, and 1D and 2D NMR data. The bioassays showed weak sterilization activities for compound 3. Compounds 1-8 were obtained from A. sampsonii for the first time.
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5.Preparation of a novel monoclonal antibody againstα-galactosidase from Bacteroides fragilis for detection of minimal residual enzyme in universal red blood cells
Subo LI ; Zhimin YUN ; Hongwei GAO ; Xue ZHANG ; Yingxia TAN ; Shikun ZHANG ; Shouping JI ; Feng GONG
Military Medical Sciences 2015;(4):302-305
Objective To establish a method of quantiying trace α-galactosidase from Bacteroides fragilis in enzymatic conversion of blood group B to O red blood cells ( B-ECO RBCs) .Methods BALB/c mice were immunized with purified recombinant B.fragilisα-galactosidase ( the purity>90%) to prepare monoclonal antibodies.The ascites were prepared using hybridoma cell lines stably secreting antibody and purified by HiTrap rProtein A column.The antibody titer and spe-cificity were detected by ELISA and Western blotting, respectively.Purified monoclonal antibody and rabbit polyclonal an-tibody were applied to detect residual enzyme in B-ECO RBCs and the washing solution was analyzed by indirect ELISA. Results A high titer and purity antibody was obtained.Western blotting showed that the antibody specifically reacted with B.fragilisα-galactosidase.Moreover, indirect ELISA was sensitive enough to detect the minimal amount of residualα-gal-actosidase at the concentration of 1 ng/ml.After four repeat washing cycles with 1∶4 ( v/v) phosphate-buffered saline, the amount of residual enzyme in B-ECO RBCs was less than 10 ng/ml.Conclusion An effective method of detecting the min-imal amount of residual α-galactosidase in blood conversion is established for safety evaluation of universal RBCs prepara-tion by enzymatic treatment.
6.Correlation of Serum Pepsinogens and Gastrin 17 With Age in Patients with Chronic Atrophic Gastritis
Huiling GUO ; Guangzhou GAO ; Jinzhuo ZHANG ; Yingxia HAO
Chinese Journal of Gastroenterology 2018;23(3):169-172
Background:Chronic atrophic gastritis(CAG)is a common disease of digestive system and is a precancerous lesion of the intestinal type gastric cancer. Serum pepsinogens(PGs)and gastrin 17(G17)are biological markers of gastric mucosal lesions,which have a prominent role in diagnosis of CAG and screening of early gastric cancer. Aims:To study the correlation of serum PGs and G17 with age in patients with CAG. Methods:A total of 582 CAG patients admitted from Jan. 2016 to Sep. 2017 at the Baoding First Central Hospital were enrolled. The levels of serum PGⅠ,PGⅡ and G17 were determined by ELISA,and the PGⅠ/ PGⅡ ratio(PGR)was calculated. The correlations of these indices with the clinical data of CAG patients were analyzed. Results:The levels of serum PGⅠ and PGⅡ were increased with age(P<0.01),and the levels of PGR and G17 were decreased with age(P <0.05). Spearman rank correlation coefficient analysis showed that the levels of PGⅠ and PGⅡ were positively correlated with age(rs=0.374,P<0.01;rs=0.559, P<0.01),and the levels of PGR and G17 were negatively correlated with age(rs= -0.649,P<0.01;rs= -0.141, P<0.05). Conclusions:The levels of serum PGⅠ,PGⅡand G17 in patients with CAG were correlated with age. When serum PGs and G17 are used as serological indicators for diagnosis of CAG and screening of early gastric cancer,the impact of age on these indices should be taken into account.
7.Quality control and safeguarding measures for the whole-process management of scientific research in tertiary hospitals
Ran YI ; Tianyu CAO ; Yingxia XU ; Lejie GAO ; Xueming CHEN
Chinese Journal of Medical Science Research Management 2018;31(2):158-160,封3-封4
Objective Sharing and exploring the efficient development of scientific research management with peers in large tertiary general hospital.Methods This article took the whole process management of scientific research in our hospital during the past five years as example,analyzed and elaborated the quality control and related safeguarding measures.Results According to the comparison between 2012 and 2016,the number of SCI papers published,the number of National Natural Science Funding and other important indicators all have been increased significantly in our hospital.Conclusions The implementation of the whole process quality control management and security measures of scientific research helps hospital to achieve initial and efficient development in research management.
8.Treatment of obstructive sleep apnea-hypopnea syndrome for children refractory asthma.
Yingxia LU ; Qinglong GU ; Chong PANG ; Fan GAO ; Chen LIU ; Jun DU ; Jing ZHAO ; Yuzhi CHEN
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2014;49(6):462-467
OBJECTIVEThe aim of this study was to understand the effect of different treatment of obstructive sleep apnea-hypopnea syndrome (OSAHS) for refractory asthma in children.
METHODSFifty two children (32 in surgical group, 20 in conservative group) with refractory asthma and OSAHS were included in the study. All children received asthma condition assessment and polysomnography (PSG) examination before and after treatment, and were followed up for 6 months.
RESULTSAll children got improved in PSG values 3 months after treatment, more significant improvement was achieved in surgical group than in conservative group (P < 0.05). While compared of OSAHS treatment, there were 2 cure, 6 notable effective, 9 effective, 3 in vain cases in conservative group, 8 cure, 16 notable effective, 8 effective, 0 in vain cases in surgery group. There was significant difference between the two groups (χ² = 8.91, P = 0.031). All children got improved in asthma condition evaluation parameters and decreased the use number of short acting β2 agonists after 6 months treatment. More significant improvement was achieved in surgical group than in conservative group. The differences of all the items had statistical significance (P < 0.05). There was statistical correlation between days mutation rate of peak expiratory flow (PEF) and apnea hypopnea index (r = 0.712, P < 0.01), and between days mutation rate of PEF and lowest oxygen saturation (r = 0.726, P < 0.01).
CONCLUSIONSActive treatment of OSAHS can improve asthma symptoms and reduce asthma medication effectively. The curative effect of surgical treatment is superior to conservative treatment.
Adenoidectomy ; Asthma ; complications ; drug therapy ; surgery ; Child ; Child, Preschool ; Female ; Humans ; Male ; Sleep Apnea, Obstructive ; complications ; drug therapy ; surgery ; Tonsillectomy
9.Clinical efficacy and safety of omalizumab in the treatment of chronic spontaneous urticaria
Yu ZHANG ; Yingxia GAO ; Ningyan GU ; Hong ZHU ; Jingjing CHEN ; Qingjie HU ; Min ZHOU ; Yuanyuan DUAN ; Xu YAO
Chinese Journal of Dermatology 2021;54(7):582-585
Objective:To evaluate the clinical efficacy and safety of omalizumab in the treatment of chronic spontaneous urticaria (CSU) .Methods:Clinical data were collected from 60 patients, who were diagnosed with CSU and received subcutaneous injections of omalizumab at a dose of 300 mg once every 4 weeks for 3 sessions in Hospital of Dermatology, Chinese Academy of Medical Sciences and Peking Union Medical College from March 2020 to September 2020, and retrospectively analyzed. At weeks 0, 2, 4, 6, 8, 10 and 12, urticaria activity score over 7 days (UAS7) and chronic urticaria quality of life (CU-Q2oL) score were used to evaluate clinical symptoms and quality of life of patients. Changes in the use of other drugs were evaluated before and after the treatment with omalizumab. Paired t test was used to compare UAS7 or CU-Q2oL score before and after treatment. Results:All the 60 CSU patients received 12 weeks of omalizumab treatment. The baseline UAS7 score was 22.37 ± 8.88 points; after one session of the treatment, the UAS7 score dropped to 2.01 ± 5.13 points, reaching the treatment plateau; at week 12, it dropped to 0.6 ± 2.63 points, and 0 point (complete control) in 93.3% of the patients, 1-6 points (favorable control) in 3.3%; the time required for UAS7 score to decrease to 0 point was 22.4 ± 3.2 days. The baseline CU-Q2oL score was 34.10 ± 15.01 points; after one session of the treatment, the CU-Q2oL score dropped to 2.41 ± 7.18 points, reaching the treatment plateau; at week 12, it was 0.56 ± 2.90 points; the time required for CU-Q2oL score to drop to 0 point was 21.15 ± 16.02 days. After the combination treatment with omalizumab, a gradual decrease in dosage or withdrawal of previous therapeutic drugs was realized. At week 12, 39 patients (65%) achieved complete control, and withdrew all therapeutic drugs except omalizumab. During the treatment and follow-up, omalizumab showed good safety, and no adverse reactions were observed.Conclusion:Omalizumab at a dose of 300 mg once every 4 weeks is markedly effective and safe for the treatment of CSU, providing a new treatment option for CSU patients with poor response to traditional therapy.
10.Ribavirin is effective against drug-resistant H7N9 influenza virus infections.
Yuhai BI ; Gary WONG ; Yingxia LIU ; Lei LIU ; George F GAO ; Yi SHI
Protein & Cell 2016;7(8):611-614